AOP1.2 Antibody

Basic Research Questions

• What is AOP1.2 Antibody and what is its target protein?

AOP1.2 Antibody refers to a monoclonal antibody targeting AOP1 (Antioxidant-like Protein 1), also known as PRDX3 (Peroxiredoxin 3). This protein exists in two relevant contexts in scientific literature:

• As a 25 kDa antioxidant protein that reduces intracellular reactive oxygen species

• As albumin-associated O-glycoprotein 1 (AOP1), a 107 kDa heavily O-glycosylated protein that forms complexes with natural antibodies

The antibody recognizes epitopes specific to these proteins, with research showing AOP1/PRDX3 plays a crucial role in mitochondrial redox regulation. The ".2" designation likely indicates a specific clone or version of the antibody.

• What are the recommended protocols for using AOP1.2 Antibody in Western blotting?

For optimal Western blot results with AOP1.2 Antibody:

When blotting for AOP1/PRDX3, non-reducing conditions may better preserve epitope structure. Research indicates that this antibody can detect AOP1 in multiple species including human, mouse, rat, hamster, monkey, chicken, and canine samples . For albumin-associated AOP1, bands at higher molecular weights may be observed due to protein complexes.

• How should AOP1.2 Antibody be stored and handled to maintain its efficacy?

Proper storage and handling are critical for maintaining antibody activity:

Research on antibody stability indicates:

• Freeze-thaw cycles significantly reduce antibody activity; limit to ≤5 cycles

• Adding equal volume of glycerol as cryoprotectant before freezing enhances stability

• For concentrate products, centrifuge briefly before opening to collect solution

A study examining monoclonal antibody stability demonstrated that proper aliquoting can extend functional lifespan by preventing aggregation and maintaining epitope recognition efficiency.

• What cross-reactivity considerations are important when using AOP1.2 Antibody across species?

Understanding cross-species reactivity is essential for comparative studies:

When planning cross-species experiments:

• Always validate antibody specificity in each new species

• Consider epitope conservation through sequence alignment analysis

• Include appropriate positive controls from validated species

• Adjust antibody concentration based on species-specific optimization

Research demonstrates that cross-reactivity predictions based on sequence homology alone are insufficient; functional validation is essential for each new species application.

• How can researchers validate AOP1.2 Antibody specificity in experimental systems?

A comprehensive validation strategy includes:

Research findings emphasize that when validating antibodies:

• Genetic approaches provide the most definitive validation

• At least two independent validation methods should be employed

• Validation should be performed for each new application or cell/tissue type

• Positive and negative controls should be run in parallel

Studies of conformation-dependent antibodies highlight the importance of validation across multiple experimental conditions, as epitope recognition can vary dramatically based on protein folding states .

Advanced Research Questions

• How can AOP1.2 Antibody be used to investigate oxidative stress mechanisms in disease models?

AOP1/PRDX3 antibodies enable detailed investigation of redox biology in multiple disease contexts:

Research demonstrates that AOP1/PRDX3 shows altered expression in various pathological conditions. For instance, age-dependent increases in AOP1 expression have been documented in the hippocampus, suggesting its involvement in age-related oxidative stress responses .

Methodology should include:

• Parallel assessment of oxidative damage markers (8-OHdG, protein carbonylation)

• Measurement of antioxidant system components (SOD, catalase)

• Analysis of mitochondrial function parameters

• Correlation with clinical or behavioral outcomes

• What are the technical considerations for using AOP1.2 Antibody in co-immunoprecipitation studies?

Optimizing co-immunoprecipitation with AOP1.2 Antibody requires careful attention to:

Research on albumin-associated O-glycoproteins (AOP1/AOP2) demonstrates that these proteins form complexes with anti-α-galactoside and anti-β-glucoside antibodies , highlighting the importance of buffer conditions that preserve native protein interactions.

When investigating protein complexes:

• Use gentle lysis conditions to maintain physiological interactions

• Consider crosslinking approaches for transient interactions

• Include appropriate controls (IgG, pre-immune serum)

• Validate interactions through reciprocal co-IP or alternative methods

• How does epitope recognition by AOP1.2 Antibody vary across different protein conformational states?

Protein conformation significantly impacts antibody recognition:

Research on conformation-dependent antibodies demonstrates that epitope recognition can be highly specific to particular protein states. Studies with fibril-specific antibodies have shown they recognize distinct assembly states of proteins while failing to detect other conformational variants .

For redox-sensitive proteins like PRDX3:

• The reduced and oxidized forms may present different epitopes

• Catalytic cycle intermediates might affect antibody recognition

• Oligomerization state can influence epitope accessibility

• Post-translational modifications may mask or reveal epitopes

• How can AOP1.2 Antibody be applied in studying autoimmune responses?

Antibodies against AOP1 can provide insights into autoimmune mechanisms:

Recent research has demonstrated intriguing relationships between viral infections and autoantibody production. Studies show that SARS-CoV-2 infection can elicit autoantibodies against apolipoprotein A-1 (AAA1), which predict COVID-19 symptom persistence . This model could be applied to investigate potential autoimmune responses against AOP1.

When studying autoimmune phenomena:

• Include healthy control samples

• Consider temporal dynamics of antibody responses

• Correlate antibody levels with disease activity markers

• Examine epitope spreading phenomena

• What methodological approaches can improve AOP1.2 Antibody performance in immunohistochemistry and immunofluorescence?

Optimizing immunostaining protocols requires systematic refinement:

Research on monoclonal antibodies in tissue staining demonstrates the critical importance of proper controls:

• Positive control tissues with known target expression

• Negative controls using isotype-matched irrelevant antibodies

• Absorption controls using immunizing peptide

• Genetic knockout tissues when available

For AOP1/PRDX3 detection:

• Mitochondrial markers can confirm subcellular localization

• Oxidative stress models can validate functional relevance

• Co-staining with cell-type specific markers enables population analysis

• 3D confocal imaging may resolve submitochondrial distribution

Studies using conformation-dependent antibodies highlight the importance of fixation method selection, as different fixatives can dramatically alter epitope accessibility and recognition .

Applied Research Case Study: AOP1 Antibodies in Translational Research

A recent study examining age-dependent changes in AOP1/PRDX3 expression demonstrated significant upregulation in aged hippocampal tissue compared to young controls, suggesting enhanced antioxidant defense mechanisms with aging . Researchers used a combination of Western blotting (0.25 μg/mL antibody concentration) and immunohistochemistry (2.5 μg/mL) to characterize both total protein levels and spatial distribution within brain regions.